乳杆菌微小膜蛋白对肠上皮细胞Caco-2的细胞毒性研究  

Cytotoxicity of micro integral membrane protein against intestinal epithelial Caco-2 cells

作　　者：屈潇 尹明明[1] 沈通一[2,3] 严雪冰[2,3] 秦环龙[2,3] QU Xiao;YIN Mingming;SHEN Tongyi;YAN Xuebing;QIN Huanlong(Shanghai Clinical College of Anhui Medical University,Shanghai 200072,China)

机构地区：[1]安徽医科大学上海临床学院,上海200072 [2]同济大学附属第十人民医院,上海200072 [3]同济大学医学院肠道疾病研究所,上海200072

出　　处：《中国微生态学杂志》2018年第7期745-749,共5页Chinese Journal of Microecology

基　　金：国家自然科学基金(81472262;81472262;81070293);2017年度黎介寿肠道屏障研究专项基金(LJS201701)

摘　　要：目的利用脂多糖(lipopolysaccharide,LPS)刺激模拟体外炎症环境,观察不同浓度下乳杆菌微小膜蛋白(micro integral membrane protein,MIMP)对肠上皮细胞Caco-2的生物学影响,评估其细胞毒性作用。方法首先通过CCK-8实验检测在LPS刺激后不同浓度MIMP(0.01、0.1和1ng/mL)对Caco-2细胞增殖活性的影响,并利用Toll样受体4(Toll-like receptor 4,TLR4)抑制剂作为阳性对照。其次利用流式细胞术检测不同浓度下MIMP对Caco-2细胞凋亡及细胞周期的影响。结果在12h的特定孵育时间内,单独应用不同浓度MIMP及TLR4抑制剂对Caco-2细胞增殖活性无显著影响(P>0.05),但是MIMP可以拮抗LPS对Caco-2细胞的促增殖作用(P<0.05)。不同浓度的MIMP对Caco-2细胞的周期和凋亡无明显影响(P>0.05)。结论不同浓度MIMP对Caco-2细胞的增殖、凋亡及细胞周期无明显影响,并可以拮抗LPS的促细胞增殖作用,因此具有较高的安全性,有望用于炎症性肠病的治疗。Objective To evaluate the biological effects of micro integral membrane protein（MIMP）on intestinal epithelial cells Caco-2 and its cytotoxicity under different concentrations.Methods Firstly,Caco-2 cells were stimulated by using LPS,the proliferation of which was detected with CCK-8 assay under different concentrations of MIMP（0.01,0.1 and 1 ng/mL）,with toll-like receptor 4（TLR4）inhibitors as the positive control.Secondly,flow cytometry was used to observe the impact of MIMP on the cell cycle and apoptosis of Caco-2 cells.Results Within specific incubation time of 12 hours,different concentrations of MIMP or TLR4 inhibitors alone had no significant effects on the proliferation of Caco-2 cells（P〈0.05）.However,MIMP antagonized the effect of LPS on the proliferation of Caco-2 cells（P〈0.05）.Different concentrations of MIMP had no significant influence on the cycle and apoptosis of Caco-2 cells,either（P〈0.05）.Conclusion MIMP can antagonize the effect of LPS on cell proliferation without obvious impact on the cell cycle,proliferation of Caco-2 cells.Therefore,MIMP may have a high safety in the clinical treatment of IBD.

关 键 词：脂多糖 微小膜蛋白 CACO-2细胞 炎症性肠病 肠道细胞功能 

分 类 号：R574[医药卫生—消化系统]